title: Chemical and structural insights into the regioversatility of the aminoglycoside acetyltransferase Eis.
abstract: A recently discovered cause of tuberculosis resistance to a drug of last resort, the aminoglycoside kanamycin, results from modification of this drug by the enhanced intracellular survival (Eis) protein. Eis is a structurally and functionally unique acetyltransferase with an unusual capability of acetylating aminoglycosides at multiple positions. The extent of this regioversatility and its defining protein features are unclear. Herein, we determined the positions and order of acetylation of five aminoglycosides by NMR spectroscopy. This analysis revealed unprecedented acetylation of the 3''-amine of kanamycin, amikacin, and tobramycin, and the γ-amine of the 4-amino-2-hydroxybutyryl group of amikacin. A crystal structure of Eis in complex with coenzyme A and tobramycin revealed how tobramycin can be accommodated in the Eis active site in two binding modes, consistent with its diacetylation. These studies, describing chemical and structural details of acetylation, will guide future efforts towards designing aminoglycosides and Eis inhibitors to overcome resistance in tuberculosis.
authors.affiliation: Department of Medicinal Chemistry, University of Michigan, Ann Arbor, MI 48109 (USA); Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109 (USA).
authors.forename: Jacob L
authors.initials: JL
authors.lastname: Houghton
authors.forename: Tapan
authors.initials: T
authors.lastname: Biswas
authors.forename: Wenjing
authors.initials: W
authors.lastname: Chen
authors.forename: Oleg V
authors.initials: OV
authors.lastname: Tsodikov
authors.forename: Sylvie
authors.initials: S
authors.lastname: Garneau-Tsodikova
mesh_terms: D000107:Acetylation; D000123:Acetyltransferases / Q000737:chemistry* / Q000378:metabolism; D000583:Amikacin / Q000737:chemistry / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D020134:Catalytic Domain; D003065:Coenzyme A / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D024881:Drug Resistance, Bacterial; D007612:Kanamycin / Q000737:chemistry / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D009169:Mycobacterium tuberculosis / Q000201:enzymology; D011485:Protein Binding; D013237:Stereoisomerism; D014031:Tobramycin / Q000737:chemistry / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D007612:Kanamycin; D000583:Amikacin; D000123:Acetyltransferases; C019681:aminoglycoside acetyltransferase; D003065:Coenzyme A; D014031:Tobramycin
keywords: bacterial resistance; enhanced intracellular survival protein; enzymatic reaction; multi-acetylation; tuberculosis